Recombinant production enables lot-to-lot consistency and is animal-cruelty-free
Western blot analysis of Phospho-Raf1(S621) on human skeletal muscle tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
In skeletal muscle, isoform 1 is more abundant than isoform 2.
Phosphorylation at Thr-269, Ser-338, Tyr-341, Thr-491 and Ser-494 results in its activation. Phosphorylation at Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2 results in its inactivation. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity. Phosphorylation at Ser-338 by PAK1 and PAK5 and Ser-339 by PAK1 is required for its mitochondrial localization. Phosphorylation at Ser-621 in response to growth factor treatment stabilizes the protein, possibly by preventing proteasomal degradation. Phosphorylation at Ser-289, Ser-296, Ser-301, Ser-338 and Ser-621 are somehow linked to the methylation potential of cells. Treatment of cells with HGF in the presence of the methylation inhibitor 5'-methylthioadenosine (MTA) results in increased phosphorylation at Ser-338 and Ser-621 and decreased phosphorylation at Ser-296, Ser-301 and Ser-338. Dephosphorylation at Ser-338 by PPP5C results in an activity decrease.; Methylated at Arg-563 in response to EGF treatment. This modification leads to destabilization of the protein, possibly through proteasomal degradation.
Cytoplasm, Cell membrane, Mitochondrion, Nucleus.
Raf-1 is a ubiquitously expressed cytoplasmic protein with intrinsic serine/threonine kinase activity. Raf-1, or c-Raf, is the cellular homolog of v-Raf, the product of the transforming gene of the 3611 strain of murine sarcoma virus. The unregulated kinase activity of the v-Raf protein is associated with cellular transformation and mitogenesis. Raf-1 is normally suppressed by its regulatory N-terminal domain. Raf-1 is activated in response to a variety of tyrosine kinase receptors as well as in response to pp60v-Src expression. Specifically, Raf-1 is phosphorylated in the catalytic domain at Ser 338 and, to a lesser extent, Ser 339. This phosphorylation requires the co-activation of PI 3-kinase and the Ras signaling pathway. Raf-1 is also phosphorylated on Tyr 340 and 341, which induces the phosphorylation of MEK. Phosphorylation of Ser 621 is essential for the catalytic activity of Raf-1 and downregulation by c-AMP-dependent protein kinase A (PKA). PKA also phosphorylates Raf-1 on Ser 43 and Ser 259. PKA phosphorylation of Ser 259 inhibits Raf-1 and decreases the phosphorylation necessary for Raf-1 activation at Ser 338.
Just like the interactions between antigens and antibodies, the higher the affinity between you and us the better.